The aim of this study is to screen source of cysteine proteinase from some vegetables and fruits. The fruit sample showed positive result on AZCL-casein plate 17 from 42 samples and vegetable showed 39 from 73 samples. The enzyme from fruit and vegetable can be classified into two types proteinase; serine and cysteine proteinases by using specific inhibitors. In this study we focus in ficain from Ficus hispida L. because it has been a very few studies. Ficain (EC3.4.22.3), cysteine proteinase from latex of Ficus hispida was purified by 80% saturated ammonium sulphate precipitation and single-step Thiopropyl Sepharose 6B covalent chromatography. The enzyme activity was completely inhibited by 2,2 dipyridyl disulphide (2PDS) and iodoacetamide, indicating that the purified enzyme is a member of cysteine proteinase. The molecular weight of purified enzyme was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The cysteine proteinase from Ficus hispida had four major bands with molecular weight range of 23,000 to 45,000 dalton. The purified multiple form enzymes were glycoprotein because each band showed positive with GelCode® glycoprotein staining kit. Major band from SDS-PAGE at 23,000 dalton was determined for amino acid residue by Liquid Chromatography-Mass Spectrometry (LC-MS/MS). The major band of purified enzyme had 55 amino acids matched to Chain A from Carica papaya chymopapain . The three-dimensional structure of chymopapain was generated by Discovery Studio® Visualizer Version 2.01, AccelrysSoftware. A comparison of the short amino acid sequences around the catalytic sites of the chymopapain indicates a high degree of similarity. In investigating the ficain from Ficus hispida L. can rapidly catalyze the hydrolysis of Gyl-Arg-p-nitroanilide dihydroxychloride, N-Benzoyl-L-Arginine-p-nitroanilide hydrochloride, Ala-Ala-Phe-p-nitroanilide, N-succinyl-Gly-Gly-Phe-p-nitroanilide, and N-succinyl-L-Phenylalanine-p-nitroanilide. This result indicated that P1 subsite of ficain specifics for amino acid arginine.